Screening and identification for cDNA of differentially expressed genes in human primary hepatocellular carcinoma.
- Author:
Jing LI
1
;
Ben-li HAN
;
Gui-jun HUANG
;
Gui-sheng QIAN
;
Ping LIANG
;
Tong-han YANG
;
Jie CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Carcinoma, Hepatocellular; genetics; Cloning, Molecular; DNA, Complementary; chemistry; genetics; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gene Library; Humans; Liver Neoplasms; genetics; Molecular Sequence Data; Sequence Analysis, DNA
- From: Chinese Journal of Medical Genetics 2003;20(1):49-52
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEScreening and identification of differentially expressed genes in human primary hepatocellular carcinoma(HCC).
METHODSThe differentially expressed genes subtracted cDNA library of HCC constructed by suppression subtractive hybridization(SSH) technique was screened by colony in situ hybridization, then the positive clones were further screened with PCR amplification. The positive clones were sequenced and analyzed for homology in the Genbank databases with Basic Local Alignment Search Tool BLAST . The novel cDNA sequences were analyzed by Northern blot analysis.
RESULTSThirteen positive clones were obtained, and 11 cDNA sequences were identified. Sequences of 11 cDNA showed that 6 cDNA were homologous with the genes published in Genbank and 5 cDNA were unknown genes. Northern blot indicated that 3 novel cDNA(>300 bp) were only expressed in HCC.
CONCLUSIONThe subtracted cDNA library constructed by SSH technique contains differentially expressed genes of HCC. Three novel cDNA sequences might be differentially expressed genes of HCC. Further screening the library and gaining the whole gene sequence may lay a foundation for identifying differentially expressed genes in HCC.
